Studies of the mechanism of TAT-tCNTF on cell injury caused by cytochalasin D

ZHANG Hong; ZHANG Qin; LIU Ze-yuan; XU Ya-fei; QU Heng-yan

Abstract：Objective:To investigate the mechanism of TAT-tCNTF on human umbilical vein endothelial cells (HUVECs) injured by cytochalasin D (CD). Methods:After HUVECs were treated with CD and TAT-tCNTF, cell viability was determined by CCK-8. The structural changes of F-actin stained by rhodamine labeled ghost cyclic peptide were observed through the lfuorescence microscope. Concentrations of intracellular free calcium by Fluo 4-AM loading were detected through the lfuorescence microscope and lfow cytometry. Western blot was used to detect the expression of F-actin protein. Results:CD (10μmol·L-1) reduced the cell viability of HUVECs (P< 0.000 1), while TAT-tCNTF (10 μg·mL-1 ) significantly increased the cell viability of HUVECs (P=0.003). F-actin with CD was fractured and reduced, while it was improved after subsequent treatment with TAT-tCNTF. TAT-tCNTF alleviated the calcium overload caused by CD. Flow cytometry showed mean lfuorescence intensity (MFI) of Ca2+in CD group was stronger than that in control group, the MFI of Ca2+in (CD+TAT-tCNTF) group was lower than that in CD group. Western blot showed the expression of F-actin protein had almost no change among all groups. Conclusion:TAT-tCNTF ameliorated the cell injury caused by CD. The mechanism was associated with maintaining actin cytoskeleton and alleviating the calcium overload.